This page should point you to the many different general and lab-specific protocols describing tissue and cell lysis and serve as forum for comparison.

+

This page should point you to the many different general and lab-specific protocols describing tissue and cell lysis and serve as forum for comparison. The ups and downs of various methods from sonication, homogenization, freeze-thaw cycles, and detergent-based lysis are examined below.

== Comparison of lysis methods ==

== Comparison of lysis methods ==

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* problem: heat build up which can denature proteins (proportionate to length of sonication)

* problem: heat build up which can denature proteins (proportionate to length of sonication)

* precaution: do on ice and sonicate intermittantly

* precaution: do on ice and sonicate intermittantly

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material: ultrasonication bath or rods

=== Homogenisation ===

=== Homogenisation ===

* best for animal tissue; less suitable for cells

* best for animal tissue; less suitable for cells

* precaution: do on ice to reduce heat build-up and denaturation

* precaution: do on ice to reduce heat build-up and denaturation

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material: drills (polytron), pestle/tube (dounce) bead beaters

=== Freeze-thaw ===

=== Freeze-thaw ===

* least effective method

* least effective method

* plus: does not denature proteins as much as other methods

* plus: does not denature proteins as much as other methods

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material: pestle and mortar, liquid nitrogen

=== Detergents ===

=== Detergents ===

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* problems: detergent may inhibit subsequent reactions

* problems: detergent may inhibit subsequent reactions

* problems: detergent may disrupts protein interactions

* problems: detergent may disrupts protein interactions

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no additional material required, just the chemicals and typical tubes

== Specific protocols ==

== Specific protocols ==

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{| {{table}}

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{| {{sorttable}}

! style="background:lightgrey"|description/link

! style="background:lightgrey"|description/link

! style="background:lightgrey"|target

! style="background:lightgrey"|target

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| protein

| protein

| detergent (Triton)

| detergent (Triton)

−

| eukaryotic cells

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| cells

|-

|-

| [[Sauer:Lysing E. coli with Lysozymes]]

| [[Sauer:Lysing E. coli with Lysozymes]]

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| protein

| protein

| detergent (RIPA buffer)

| detergent (RIPA buffer)

−

| cell line

+

| cells

|-

|-

| [[Streptomyces:Protocols/Mini-Maxi Prep]]

| [[Streptomyces:Protocols/Mini-Maxi Prep]]

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| protein

| protein

| detergent (SDS, NP40)

| detergent (SDS, NP40)

−

| cell line

+

| cells

|-

|-

| [[Eccles:Protein Lysates from Tissue]]

| [[Eccles:Protein Lysates from Tissue]]

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| RNA

| RNA

| phenol

| phenol

−

| cell line

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| cells

+

|-

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| [[Sauer:RNA Purification from E. coli]]

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| RNA

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| various options

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| bacteria

|-

|-

| add another method's name

| add another method's name

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| type of starting material

| type of starting material

|}

|}

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== Related hub pages ==

== Related hub pages ==

Revision as of 02:45, 19 December 2008

This page should point you to the many different general and lab-specific protocols describing tissue and cell lysis and serve as forum for comparison. The ups and downs of various methods from sonication, homogenization, freeze-thaw cycles, and detergent-based lysis are examined below.